Human OA chondrocytes were transfected with anti‐miRNAs (50 nM; Ambion, Foster City, CA, USA or Qiagen, Hilden, Germany) at a 50 nM concentration, using the calcium phosphate precipitation method 27 . Following transfection, chondrocytes were pre‐treated with EGCG (20–50 μM) and then stimulated with IL‐1β (5 ng/ml) for 8–24 hrs to analyse the expression of miRNA, mRNA or protein.
Anti mirnas
Manufactured by Thermo Fisher Scientific
Sourced in United States
Anti-miRNAs are synthetic oligonucleotides designed to inhibit the function of specific microRNAs (miRNAs) in biological systems. They serve as valuable tools for studying the role of miRNAs in cellular processes and disease mechanisms.
Automatically generated - may contain errors
Lab products found in correlation
Hiperfect transfection reagent, by Qiagen (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Pdgfr β, by Santa Cruz Biotechnology (1 mentions)
Anti rb, by Cell Signaling Technology (1 mentions)
Cyclin a, by Santa Cruz Biotechnology (1 mentions)
Fabp4, by Cell Signaling Technology (1 mentions)
Pdgfrβ, by Horizon Discovery (1 mentions)
Pparγ, by Cell Signaling Technology (1 mentions)
Vinculin, by Santa Cruz Biotechnology (1 mentions)
Cp 673451, by Selleck Chemicals (1 mentions)
Icg 001, by Selleck Chemicals (1 mentions)
C ebpα, by Cell Signaling Technology (1 mentions)
P rb ser780, by Cell Signaling Technology (1 mentions)
Smad4, by Santa Cruz Biotechnology (1 mentions)
Cyclin d1, by Santa Cruz Biotechnology (1 mentions)
4 protocols using anti mirnas
1
Chondrocyte Regulation by miRNA and EGCG
2
MCF-7 Cell Transfection and AuNP Exposure
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MCF-7 cell transfection was carried out with miRNA inhibitors (anti-miRNAs; 100 nM; Ambion/Qiagen). HiPerfect Transfection Reagent was used according to the manufacturer’s instructions (Qiagen, Germantown, MD, USA). After 72 h post-transfection, MCF-7 cells were treated with 0.2–0.6 µg/mL of AuNPs for 2 h or stimulated with PMA (0.1 µM) for 24 h, and cell lysate or RNA samples were prepared for further analysis.
3
Transfection of Pre- and Anti-miRNAs
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Pre- and anti-miRNAs were purchased from Ambion (Austin, TX, USA). Transfections were performed with Lipofectamine (RNAimax; Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. Transfection complexes were prepared with 50 to 100 nM pre- or anti-miRNA, and cells were transfected 24 hours after plating.
4
Antibody and Small Molecule Reagents Protocol
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The sources of antibodies were as follows: PDGFRΒ (polyclonal, catalog number sc-432), vinculin (monoclonal, sc-73614), cyclin D1 (monoclonal, sc-8396), and cyclin A (polyclonal, sc-751) from Santa Cruz Biotechnology; SMAD4 (monoclonal, #38454, raised against peptide corresponding to residues surrounding D165), YAP1 (polyclonal, #4912), C/EBPα (polyclonal, #2295), FABP4 (polyclonal, #2120), PPARγ (#2430, raised against peptide corresponding to residues surrounding D69), and p-RB (Ser780) (polyclonal, #9307, raised against phosphopeptide corresponding to residues surrounding S780) from Cell Signaling Technology; and anti-RB (polyclonal, 09–100, raised against linear protein corresponding to human RB-like protein 2 at and around the C-terminus) from EMD Millipore. miRNAs (nonspecific miRNA, miR-193b) and antimiRNAs (nonspecific antimiRNA, anti-miR-193b) were purchased from Ambion. Smartpool siRNAs for SMAD4, PDGFRΒ and YAP1 were purchased from Dharmacon. CP-673451 and ICG-001 were purchased from Selleck Chemicals.
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